TY - JOUR

T1 - Simplifying Barley Leaf Rust Research

T2 - An Easy and Reproducible Infection Protocol for Puccinia hordei on a Small Laboratory Scale

AU - Skoppek, Caroline I.

AU - Streubel, Jana

N1 - Funding Information: This work was funded by university core funding only. We thank Gwendolin Wehner and Frank Ordon from the Julius Kühn Institute in Quedlinburg for Ph isolate I-80. We thank Jens Boch for general support. This protocol was derived from Skoppek et al. (2022). The staining method was adapted for barley from Redkar et al. (2018).

PY - 2023/7/20

Y1 - 2023/7/20

N2 - Barley (Hordeum vulgare) is one of the most important agricultural crops in the world, but pathogen infections regularly limit its annual yield. A major threat is the infection with the biotrophic leaf rust fungus, Puccinia hordei. Rust fungi have a complex life cycle, and existing resistances can be easily overcome. To address this problem, it is crucial to develop barley varieties with improved and durable resistance mechanisms. An essential step towards this goal is a simple and reproducible infection protocol to evaluate potential resistance phenotypes in the lab. However, available protocols sometimes lack detailed procedure or equipment information, use spore application methods that are not suitable for uniform spore dispersion, or require special mineral oils or engineered fluids. In addition, they are often optimized for pathogen-dedicated greenhouses or phytochambers, which may not be available to every research institute. Here, we describe an easy and user-friendly procedure to infect barley with Puccinia hordei on a small laboratory scale. This procedure utilizes inexpensive and simple tools to evenly split and apply spores to barley leaves. The treated plants are incubated in affordable and small phytocabinets. Our protocol enables a quick and reproducible infection of barley with leaf rust, a method that can easily be transferred to other rust fungi, including stripe rust, or to other plant species.

AB - Barley (Hordeum vulgare) is one of the most important agricultural crops in the world, but pathogen infections regularly limit its annual yield. A major threat is the infection with the biotrophic leaf rust fungus, Puccinia hordei. Rust fungi have a complex life cycle, and existing resistances can be easily overcome. To address this problem, it is crucial to develop barley varieties with improved and durable resistance mechanisms. An essential step towards this goal is a simple and reproducible infection protocol to evaluate potential resistance phenotypes in the lab. However, available protocols sometimes lack detailed procedure or equipment information, use spore application methods that are not suitable for uniform spore dispersion, or require special mineral oils or engineered fluids. In addition, they are often optimized for pathogen-dedicated greenhouses or phytochambers, which may not be available to every research institute. Here, we describe an easy and user-friendly procedure to infect barley with Puccinia hordei on a small laboratory scale. This procedure utilizes inexpensive and simple tools to evenly split and apply spores to barley leaves. The treated plants are incubated in affordable and small phytocabinets. Our protocol enables a quick and reproducible infection of barley with leaf rust, a method that can easily be transferred to other rust fungi, including stripe rust, or to other plant species.

KW - Barley

KW - Biotrophic fungi

KW - Golden Promise

KW - Puccinia hordei

KW - Resistance assessment

KW - Rust infection

KW - Wheat germ agglutinin Alexa Fluor 488 staining

UR - http://www.scopus.com/inward/record.url?scp=85166552380&partnerID=8YFLogxK

U2 - 10.21769/BioProtoc.4721

DO - 10.21769/BioProtoc.4721

M3 - Article

AN - SCOPUS:85166552380

VL - 13

JO - Bio-protocol

JF - Bio-protocol

IS - 14

M1 - e4721

ER -